Stable pharmaceutical composition and methods of using same

ABSTRACT

The present invention relates to, inter alia, pharmaceutical compositions comprising a polyunsaturated fatty acid and to methods of using the same to treat or prevent cardiovascular-related diseases.

PRIORITY CLAIM

This application claims priority from U.S. application Ser. No.12/769,885 filed Apr. 29, 2010 which claims priority to U.S. provisionalpatent application 61/173,763, filed Apr. 29, 2009, the entireties ofeach of which are hereby incorporated by reference herein.

BACKGROUND

Mixed omega-3 fatty acid esters are typically encapsulated in type 2agelatin capsules containing gelatin (˜43.4%), glycerol (˜20%) and water(˜36.6%) and do not experience stability problems throughout their shelflife. While chemically modified gelatins such as succinated/succinylatedgelatin have been used to encapsulate reactive fill ingredients, suchgelatin is not approved for use in the U.S. and other markets.

SUMMARY

We have unexpectedly found that high purity eicosapentaenoic acid (EPA)is more susceptible to oxidative degradation than mixed omega-3-acidethyl esters. In various embodiments, the invention providespharmaceutical compositions comprising a fatty acid or a derivativethereof in a capsule shell that resists, hinders, attenuates, orprevents oxidation of the fatty acid or fatty acid derivative, forexample to a greater extent than is provided by a standard type IIacapsule shell. In a related embodiment, the fatty acid compriseseicosapentaenoic acid (EPA) or a derivative of EPA, for example ethyleicosapentaetaenoate (ethyl-EPA or F-EPA). In another embodiment, thefatty acid comprises ultra-pure EPA.

In one embodiment, the invention provides a pharmaceutical compositioncomprising ultra-pure EPA encapsulated in a capsule shell, where theultra-pure EPA has a baseline peroxide value not greater than about 5Meq/kg and upon storage of the composition at 23° C. and 50% RH for aperiod of time, that ultra-pure EPA has a second peroxide value notgreater than about 20 Meq/kg.

In other embodiments, the invention provides a pharmaceuticalcomposition comprising EPA (e.g. E-EPA or ultra pure E-EPA) encapsulatedin a capsule shell comprising a film forming material and a hygroscopicplasticizer, wherein the weight ratio of film-forming material tohygroscopic plasticizer is not less than about 2.5:1. Further, thecapsule shell can optionally comprise a non-hygroscopic plasticizer. Inone embodiment, the capsule contains no chemically modified gelatin, forexample succinated or succinylated gelatin.

In still other embodiments, the present invention provides methods oftreating or preventing a cardiovascular-related disease usingcompositions as described herein.

These and other embodiments of the present invention will be disclosedin further detail herein below.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows dissolution profile of an inventive capsule compositioncontaining ˜500 mg E-EPA versus a composition comprising EPA in asuccinated gelatin capsule.

FIG. 2 shows bioavailability of 300 mg of EPA in succinated gelatincapsules.

FIG. 3 shows bioavailability of an inventive AMR101 capsule compositioncontaining ˜500 mg E-EPA.

DETAILED DESCRIPTION

While the present invention is capable of being embodied in variousforms, the description below of several embodiments is made with theunderstanding that the present disclosure is to be considered as anexemplification of the invention, and is not intended to limit theinvention to the specific embodiments illustrated. Headings are providedfor convenience only and are not to be construed to limit the inventionin any manner. Embodiments illustrated under any heading may be combinedwith embodiments illustrated under any other heading.

The use of numerical values in the various quantitative values specifiedin this application, unless expressly indicated otherwise, are stated asapproximations as though the minimum and maximum values within thestated ranges were both preceded by the word “about.” In this manner,slight variations from a stated value can be used to achievesubstantially the same results as the stated value. Also, the disclosureof ranges is intended as a continuous range including every valuebetween the minimum and maximum values recited as well as any rangesthat can be formed by such values. Also disclosed herein are any and allratios (and ranges of any such ratios) that can be formed by dividing arecited numeric value into any other recited numeric value. Accordingly,the skilled person will appreciate that many such ratios, ranges, andranges of ratios can be unambiguously derived from the numerical valuespresented herein and in all instances such ratios, ranges, and ranges ofratios represent various embodiments of the present invention.

Polyunsaturated Fatty Acids

In one embodiment, compositions of the invention comprise apolyunsaturated fatty acid as an active ingredient. In anotherembodiment, compositions of the invention comprise EPA as an activeingredient. The term “EPA” as used herein refers to eicosapentaenoicacid (e.g. eicosa-5,8,11,14,17-pentaenoic acid) and/or apharmaceutically acceptable ester, derivative, conjugate or saltthereof, or mixtures of any of the foregoing.

In one embodiment, the EPA comprises all-ciseicosa-5,8,11,14,17-pentaenoic acid. In another embodiment, the EPA isin the form of an eicosapentaenoic acid ester. In another embodiment,the EPA comprises a C₁-C₅ alkyl ester of EPA. In another embodiment, theEPA comprises eicosapentaenoic acid ethyl ester, eicosapentaenoic acidmethyl ester, eicosapentaenoic acid propyl ester, or eicosapentaenoicacid butyl ester. In still another embodiment, the EPA comprises all-ciseicosa-5,8,11,14,17-pentaenoic acid ethyl ester.

In still other embodiments, the EPA comprises ethyl-EPA, lithium EPA,mono, di- or triglyceride EPA or any other ester or salt of EPA, or thefree acid form of EPA. The EPA may also be in the form of a2-substituted derivative or other derivative which slows down its rateof oxidation but does not otherwise change its biological action to anysubstantial degree.

The term “pharmaceutically acceptable” in the present context means thatthe substance in question does not produce unacceptable toxicity to thesubject or interaction with other components of the composition.

In one embodiment, EPA present in a composition of the inventioncomprises ultra-pure EPA. The term “ultra-pure” as used herein withrespect to EPA refers to a composition comprising at least 96% by weightEPA (as the term “EPA” is defined and exemplified herein). Ultra-pureEPA can comprise even higher purity EPA, for example at least 97% byweight EPA or at least 98% by weight EPA, wherein the EPA is any form ofEPA as set forth herein. Ultra-pure EPA can further be defined (e.g.impurity profile) by any of the description of EPA provided herein.

In other embodiments, EPA is present in a composition of the inventionin an amount of about 50 mg to about 5000 mg, about 75 mg to about 2500mg, or about 100 mg to about 1000 mg, for example about 75 mg, about 100mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350mg, about 375 mg, about 400 mg, about 425 mg, about 450 mg, about 475mg, about 500 mg, about 525 mg, about 550 mg, about 575 mg, about 600mg, about 625 mg, about 650 mg, about 675 mg, about 700 mg, about 725mg, about 750 mg, about 775 mg, about 800 mg, about 825 mg, about 850mg, about 875 mg, about 900 mg, about 925 mg, about 950 mg, about 975mg, about 1000 mg, about 1025 mg, about 1050 mg, about 1075 mg, about1100 mg, about 1025 mg, about 1050 mg, about 1075 trig, about 1200 mg,about 1225 mg, about 1250 mg, about 1275 mg, about 1300 mg, about 1325mg, about 1350 mg, about 1375 mg, about 1400 mg, about 1425 mg, about1450 mg, about 1475 mg, about 1500 mg, about 1525 mg, about 1550 mg,about 1575 trig, about 1600 mg, about 1625 mg, about 1650 mg, about 1675mg, about 1700 mg, about 1725 mg, about 1750 mg, about 1775 mg, about1800 mg, about 1825 mg, about 1850 mg, about 1875 mg, about 1900 mg,about 1925 mg, about 1950 mg, about 1975 mg, about 2000 mg, about 2025mg, about 2050 mg, about 2075 mg, about 2100 mg, about 2125 mg, about2150 mg, about 2175 mg, about 2200 mg, about 2225 mg, about 2250 mg,about 2275 mg, about 2300 mg, about 2325 mg, about 2350 mg, about 2375mg, about 2400 mg, about 2425 mg, about 2450 mg, about 2475 mg, or about2500 mg.

In various embodiments, one or more antioxidants can be present in theEPA (e.g. E-EPA or ultra pure E-EPA). Non-limiting examples of suitableantioxidants include tocopherol, lecithin, citric acid and/or ascorbicacid. One or more antioxidants, if desired, are typically present in theEPA in an amount of about 0.01% to about 0.1%, by weight, or about0.025% to about 0.05%, by weight.

In one embodiment, a composition of the invention contains not more thanabout 10%, not more than about 9%, not more than about 8%, not more thanabout 7%, not more than about 6%, not more than about 5%, not more thanabout 4%, not more than about 3%, not more than about 2%, not more thanabout 1%, or not more than about 0.5%, by weight of total fatty acids,docosahexaenoic acid or derivative thereof such as E-DHA, if any. Inanother embodiment, a composition of the invention containssubstantially no docosahexaenoic acid or derivative thereof such asE-DHA. In still another embodiment, a composition of the inventioncontains no docosahexaenoic acid or E-DHA.

In another embodiment, EPA represents at least about 60%, at least about70%, at least about 80%, at least about 90%, at least about 95%, atleast about 97%, at least about 98%, at least about 99%, or 100%, byweight, of all fatty acids present in a composition of the invention.

In another embodiment, a composition of the invention contains less than30%, less than 20%, less than 10%, less than 9%, less than 8%, less than7%, less than 6%, less than 5%, less than 4%, less than 3%, less than2%, less than 1%, less than 0.5% or less than 0.25%, by weight of thetotal composition or by weight of the total fatty acid content, of anyfatty acid other than EPA, or derivative thereof. Illustrative examplesof a “fatty acid other than EPA” include linolenic acid (LA) orderivative thereof such as ethyl-linolenic acid, arachidonic acid (AA)or derivative thereof such as ethyl-AA, docosahexaenoic acid (DHA) orderivative thereof such as ethyl-DHA, alpha-linolenic acid (ALA) orderivative thereof such as ethyl-ALA, stearadonic acid (STA) orderivative thereof such as ethyl-SA, eicosatrienoic acid (ETA) orderivative thereof such as ethyl-ETA and/or docosapentaenoic acid (DPA)or derivative thereof such as ethyl-DPA.

In another embodiment, a composition of the invention has one or more ofthe following features: (a) eicosapentaenoic acid ethyl ester representsat least 96%, at least 97%, or at least 98%, by weight, of all fattyacids present in the composition; (b) the composition contains not morethan 4%, not more than 3%, or not more than 2%, by weight, of totalfatty acids other than eicosapentaenoic acid ethyl ester; (c) thecomposition contains not more than 0.6%, 0.5%, or 0.4% of any individualfatty acid other than eicosapentaenoic acid ethyl ester; (d) thecomposition has a refractive index (20° C.) of about 1 to about 2, about1.2 to about 1.8 or about 1.4 to about 1.5; (e) the composition has aspecific gravity (20° C.) of about 0.8 to about 1.0, about 0.85 to about0.95 or about 0.9 to about 0.92; (f) the composition contains not morethan 20 ppm, 15 ppm or 10 ppm heavy metals, (g) the composition containsnot more than 5 ppm, 4 ppm, 3 ppm, or 2 ppm arsenic, and/or (h) thecomposition has a peroxide value not more than 5, 4, 3, or 2 Meq/kg.

In another embodiment, a composition useful in accordance with theinvention comprises, consists essentially of or consists of at least 95%by weight ethyl eicosapentaenoate (EPA-E), about 0.2% to about 0.5% byweight ethyl octadecatetraenoate (ODTA-E), about 0.05% to about 0.25% byweight ethyl nonaecapentaenoate (NDPA-E), about 0.2% to about 0.45% byweight ethyl arachidonate (AA-E), about 0.3% to about 0.5% by weightethyl eicosatetraenoate (ETA-E), and about 0.05% to about 0.32% ethylheneicosapentaenoate (HPA-E). In another embodiment, the composition ispresent in a capsule shell. In still another embodiment, the capsuleshell contains no chemically modified gelatin.

In another embodiment, compositions useful in accordance with theinvention comprise, consist essentially of, or consist of at least 95%,96% or 97%, by weight, ethyl eicosapentaenoate, about 0.2% to about 0.5%by weight ethyl octadecatetraenoate, about 0.05% to about 0.25% byweight ethyl nonaecapentaenoate, about 0.2% to about 0.45% by weightethyl arachidonate, about 0.3% to about 0.5% by weight ethyleicosatetraenoate, and about 0.05% to about 0.32% by weight ethylheneicosapentaenoate. Optionally, the composition contains not more thanabout 0.06%, about 0.05%, or about 0.04%, by weight, DHA or derivativethere of such as ethyl-DHA. In one embodiment the composition containssubstantially no or no amount of DHA or derivative there of such asethyl-DHA. The composition further optionally comprises one or moreantioxidants (e.g. tocopherol) in an amount of not more than about 0.5%or not more than 0.05%. In another embodiment, the composition comprisesabout 0.05% to about 0.4%, for example about 0.2% by weight tocopherol.In another embodiment, about 500 mg to about 1 g of the composition isprovided in a capsule shell. In another embodiment, the capsule shellcontains no chemically modified gelatin.

In another embodiment, compositions useful in accordance with theinvention comprise, consist essentially of, or consist of at least 96%by weight ethyl eicosapentaenoate, about 0.22% to about 0.4% by weightethyl octadecatetraenoate, about 0.075% to about 0.20% by weight ethylnonaecapentaenoate, about 0.25% to about 0.40% by weight ethylarachidonate, about 0.3% to about 0.4% by weight ethyl eicosatetraenoateand about 0.075% to about 0.25% by weight ethyl heneicosapentaenoate.Optionally, the composition contains not more than about 0.06%, about0.05%, or about 0.04%, by weight, DHA or derivative there of such asethyl-DHA. In one embodiment the composition contains substantially noor no amount of DHA or derivative there of such as ethyl-DHA. Thecomposition further optionally comprises one or more antioxidants (e.g.tocopherol) in an amount of not more than about 0.5% or not more than0.05%. In another embodiment, the composition comprises about 0.05% toabout 0.4%, for example about 0.20%, by weight tocopherol. In anotherembodiment, the invention provides a dosage form comprising about 500 mgto about 1 g of the foregoing composition in a capsule shell. In oneembodiment, the dosage form is a gel- or liquid-containing capsule andis packaged in blister packages of about 1 to about 20 capsules persheet.

In another embodiment, compositions useful in accordance with theinvention comprise, consist essentially of or consist of at least 96%,97% or 98%, by weight, ethyl eicosapentaenoate, about 0.25% to about0.38% by weight ethyl octadecatetraenoate, about 0.10% to about 0.15% byweight ethyl nonaecapentaenoate, about 0.25% to about 0.35% by weightethyl arachidonate, about 0.31% to about 0.38% by weight ethyleicosatetraenoate, and about 0.08% to about 0.20% by weight ethylheneicosapentaenoate. Optionally, the composition contains not more thanabout 0.06%, about 0.05%, or about 0.04%, by weight, DHA or derivativethere of such as ethyl-DHA. In one embodiment the composition containssubstantially no or no amount of DHA or derivative there of such asethyl-DHA. The composition further optionally comprises one or moreantioxidants (e.g. tocopherol) in an amount of not more than about 0.5%or not more than 0.05%. In another embodiment, the composition comprisesabout 0.05% to about 0.4%, for example about 0.2% by weight tocopherol.In another embodiment, the invention provides a dosage form comprisingabout 500 mg to about 1 g of the foregoing composition in a capsuleshell. In another embodiment, the capsule shell contains no chemicallymodified gelatin.

In various embodiments, the invention provides a polyunsaturated fattyacid such as EPA (e.g. E-EPA or ultra pure E-EPA) encapsulated in apharmaceutical capsule shell. In one embodiment, the capsule shellresists, hinders, attenuates, or prevents oxidation of the fatty acid orfatty acid derivative. In another embodiment, the capsule shell resists,hinders, attenuates, or prevents oxidation of the polyunsaturated fattyacid or derivative to a greater extent than a standard type IIa gelatincapsule. In another embodiment, the capsule contains no chemicallymodified gelatin, for example succinated, succinylated, pthalated,carbanylated and/or phenol carbanylated gelatin.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising EPA (e.g. E-EPA or ultra pure E-EPA) encapsulatedin a capsule shell as described herein and having a baseline peroxidevalue not greater than about 10 Meq/kg, about 9 Meq/kg, about 8 Meq/kg,about 7 Meq/kg, about 6 Meq/kg, about 5 Meq/kg, about 4 Meq/kg, about 3Meq/kg or about 2 Meq/kg, wherein upon storage of the composition at 23°C. and 50% RH for a period about 1, about 2, about 3, about 4, about 5,about 6, about 7, about 8, about 9, about 10, about 11, about 12, about13, about 14, about 15, about 16, about 17, about 18, about 19, about20, about 21, about 22, about 23 or about 24 months, the ultra-pure EPAhas a second peroxide value not greater than about 25 Meq/kg, about 24Meq/kg, about 23 Meq/kg, about 22 Meq/kg, about 21 Meq/kg, about 20Meq/kg, about 19 Meq/kg, about 18 Meq/kg, about 17 Meq/kg, about 16Meq/kg, about 15 Meq/kg, about 14 Meq/kg, about 13 Meq/kg, about 12Meq/kg, about 11 Meq/kg, about 10 Meq/kg, about 9 Meq/kg, about 8Meq/kg, about 7 Meq/g, about 6 Meq/q/kg, about 5 Meq/kg, about 4 Meq/kg,about 3 Meq/kg or about 2 Meq/kg.

The “baseline peroxide value” and “second peroxide values” can bemeasured in any suitable manner, for example by using a U.S. or PhEur orJP compendial method. Typically, a plurality of encapsulated EPAcompositions are provided, each composition containing EPA having beenencapsulated at substantially the same time. A first sampling of 1 ormore capsules from the plurality is provided, the capsules are openedand peroxide value of the EPA is measured substantially immediatelythereafter, providing an average baseline peroxide value. Atsubstantially the same time, a second sampling of 1 or more capsulesfrom the plurality are provided and are placed under desired storageconditions for a desired time period. At the end of the desired timeperiod, the capsules are opened and peroxide value of the EPA ismeasured substantially immediately thereafter, providing an averagesecond peroxide value. The baseline and second peroxide values can thenbe compared. In one embodiment, the “baseline peroxide value” and“second peroxide value” are determined using a plurality of encapsulatedEPA dosage units wherein each dosage unit was encapsulated (i.e. the EPAfilled and sealed into capsules) within a same 60 day period, same 30day period, a same 20 day period, a same 10 day period, a same 5 dayperiod or a same 1 day period.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising EPA (e.g. E-EPA or ultra pure E-EPA) encapsulatedin a capsule shell as described herein and having a baseline peroxidevalue not greater than about 10 Meq/kg, about 9 Meq/kg, about 8 Meq/kg,about 7 Meq/kg, about 6 Meq/kg, about 5 Meq/kg, about 4 Meq/kg, about 3Meq/kg or about 2 Meq/kg, wherein upon storage of the composition at 25°C. and 60% RH for a period about 1, about 2, about 3, about 4, about 5,about 6, about 7, about 8, about 9, about 10, about 11, about 12, about13, about 14, about 15, about 16, about 17, about 18, about 19, about20, about 21, about 22, about 23 or about 24 months, said compositionhas a second peroxide value not greater than about 25 Meq/kg, about 24Meq/kg, about 23 Meq/kg, about 22 Meq/kg, about 21 Meq/kg, about 20Meq/kg, about 19 Meq/kg, about 18 Meq/kg, about 17 Meq/kg, about 16Meq/kg, about 15 Meq/kg, about 14 Meq/kg, about 13 Meq/kg, about 12Meq/kg, about 11 Meq/kg, about 10 Meq/kg, about 9 Meq/kg, about 8Meq/kg, about 7 Meq/kg, about 6 Meq/kg, about 5 Meq/kg, about 4 Meq/kg,about 3 Meq/kg or about 2 Meq/kg.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising EPA (e.g. E-EPA or ultra pure E-EPA) encapsulatedin a capsule shell as described herein and having a baseline peroxidevalue not greater than about 10 Meq/kg, about 9 Meq/kg, about 8 Meq/kg,about 7 Meq/kg, about 6 Meq/kg, about 5 Meq/kg, about 4 Meq/kg, about 3Meq/kg or about 2 Meq/kg, wherein upon storage of the composition at 30°C. and 65% RH for a period about 1, about 2, about 3, about 4, about 5,about 6, about 7, about 8, about 9, about 10, about 11, about 12, about13, about 14, about 15, about 16, about 17, about 18, about 19, about20, about 21, about 22, about 23 or about 24 months, said compositionhas a second peroxide value not greater than about 25 Meq/kg, about 24Meq/kg, about 23 Meq/kg, about 22 Meq/kg, about 21 Meq/kg, about 20Meq/kg, about 19 Meq/kg, about 18 Meq/kg, about 17 Meq/kg, about 16Meq/kg, about 15 Meq/kg, about 14 Meq/kg, about 13 Meq/kg, about 12Meq/kg, about 11 Meq/kg, about 10 Meq/kg, about 9 Meq/kg, about 8Meq/kg, about 7 Meq/kg, about 6 Meq/kg, about 5 Meq/kg, about 4 Meq/kg,about 3 Meq/kg or about 2 Meq/kg.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising EPA (e.g. E-EPA or ultra pure E-EPA) encapsulatedin a capsule shell as described herein and having a baseline peroxidevalue not greater than about 10 Meq/kg, about 9 Meq/kg, about 8 Meq/kg,about 7 Meq/kg, about 6 Meq/kg, about 5 Meq/kg, about 4 Meq/kg, about 3Meq/kg or about 2 Meq/kg, wherein upon storage of the composition at 40°C. and 75% RH for a period about 1, about 2, about 3, about 4, about 5,about 6, about 7, about 8, about 9, about 10, about 11, about 12, about13, about 14, about 15, about 16, about 17, about 18, about 19, about20, about 21, about 22 about 23 or about 24 months, said composition hasa second peroxide value not greater than about 25 Meq/kg, about 24Meq/kg, about 23 Meq/kg, about 22 Meq/kg, about 21 Meq/kg, about 20Meq/kg, about 19 Meq/kg, about 18 Meq/kg, about 17 Meq/kg, about 16Meq/kg, about 15 Meq/kg, about 14 Meq/kg, about 13 Meq/kg, about 12Meq/kg, about 11 Meq/kg, about 10 Meq/kg, about 9 Meq/kg, about 8Meq/kg, about 7 Meq/kg, about 6 Meq/kg, about 5 Meq/kg, about 4 Meq/kg,about 3 Meq/kg or about 2 Meq/kg.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising EPA (e.g. E-EPA or ultra pure E-EPA) encapsulatedin a capsule shell and having a baseline peroxide value not greater thanabout 10 Meq/kg, about 9 Meq/kg, about 8 Meq/kg, about 7 Meq/kg, about 6Meq/kg, about 5 Meq/kg, about 4 Meq/kg, about 3 Meq/kg or about 2Meq/kg, wherein the capsule comprises a film-forming material and aplasticizer in a weight ratio of not less than 1.75:1 and wherein uponstorage of the composition at 23° C. and 50% RH for a period about 1,about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9,about 10, about 11, about 12, about 13, about 14, about 15, about 16,about 17, about 18, about 19, about 20, about 21, about 22, about 23 orabout 24 months, said composition has a second peroxide value notgreater than about 25 Meq/kg, about 24 Meq/kg, about 23 Meq/kg, about 22Meq/kg, about 21 Meq/kg, about 20 Meq/kg, about 19 Meq/kg, about 18Meq/kg, about 17 Meq/kg, about 16 Meq/kg, about 15 Meq/kg, about 14Meq/kg, about 13 Meq/kg, about 12 Meq/kg, about 11 Meq/kg, about 10Meq/kg, about 9 Meq/kg, about 8 Meq/kg, about 7 Meq/kg, about 6 Meq/kg,about 5 Meq/kg, about 4 Meq/kg, about 3 Meq/kg or about 2 Meq/kg.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising EPA (e.g. E-EPA or ultra pure E-EPA) encapsulatedin a capsule shell and having a baseline peroxide value not greater thanabout 10 Meq/kg, about 9 Meq/kg, about 8 Meq/kg, about 7 Meq/kg, about 6Meq/kg, about 5 Meq/kg, about 4 Meq/kg, about 3 Meq/kg or about 2Meq/kg, wherein the capsule comprises a film-forming material and aplasticizer in a weight ratio of not less than 1.75:1 and wherein uponstorage of the composition at 25° C. and 60% RH for a period about 1,about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9,about 10, about 11, about 12, about 13, about 14, about 15, about 16,about 17, about 18, about 19, about 20, about 21, about 22, about 23 orabout 24 months, said composition has a second peroxide value notgreater than about 25 Meq/kg, about 24 Meq/kg, about 23 Meq/kg, about 22Meq/kg, about 21 Meq/kg, about 20 Meq/kg, about 19 Meq/kg, about 18Meq/kg, about 17 Meq/kg, about 16 Meq/kg, about 15 Meq/kg, about 14Meq/kg, about 13 Meq/kg, about 12 Meq/kg, about 11 Meq/kg, about 10Meq/kg, about 9 Meq/kg, about 8 Meq/kg, about 7 Meq/kg, about 6 Meq/kg,about 5 Meq/kg, about 4 Meq/kg, about 3 Meq/kg or about 2 Meq/kg.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising EPA (e.g. E-EPA or ultra pure E-EPA) encapsulatedin a capsule shell and having a baseline peroxide value not greater thanabout 10 Meq/kg, about 9 Meq/kg, about 8 Meq/kg, about 7 Meq/kg, about 6Meq/kg, about 5 Meq/kg, about 4 Meq/kg, about 3 Meq/kg or about 2Meq/kg, wherein the capsule comprises a film-forming material and aplasticizer in a weight ratio of not less than 1.75:1 and wherein uponstorage of the composition at 30° C. and 65% RH for a period about 1,about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9,about 10, about 11, about 12, about 13, about 14, about 15, about 16,about 17, about 18, about 19, about 20, about 21, about 22, about 23 orabout 24 months, said composition has a second peroxide value notgreater than about 25 Meq/kg, about 24 Meq/kg, about 23 Meq/kg, about 22Meq/kg, about 21 Meq/kg, about 20 Meq/kg, about 19 Meq/kg, about 18Meq/kg, about 17 Meq/kg, about 16 Meq/kg, about 15 Meq/kg, about 14Meq/kg, about 13 Meq/kg, about 12 Meq/kg, about 11 Meq/kg, about 10Meq/kg, about 9 Meq/kg, about 8 Meq/kg, about 7 Meq/kg, about 6 Meq/kg,about 5 Meq/kg, about 4 Meq/kg, about 3 Meq/kg or about 2 Meq/kg.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising EPA (e.g. E-EPA or ultra pure E-EPA) encapsulatedin a capsule shell and having a baseline peroxide value not greater thanabout 10 Meq/kg, about 9 Meq/kg, about 8 Meq/kg, about 7 Meq/kg, about 6Meq/kg, about 5 Meq/kg, about 4 Meq/kg, about 3 Meq/kg or about 2Meq/kg, wherein the capsule comprises a film-forming material and aplasticizer in a weight ratio of not less than 1.75:1 and wherein uponstorage of the composition at 40° C. and 75% RH for a period about 1,about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9,about 10, about 11, about 11, about 12, about 13, about 14, about 15,about 16, about 17, about 18, about 19, about 20, about 21, about 22,about 23 or about 24 months, said composition has a second peroxidevalue not greater than about 25 Meq/kg, about 24 Meq/kg, about 23Meq/kg, about 22 Meq/kg, about 21 Meq/kg, about 20 Meq/kg, about 19Meq/kg, about 18 Meq/kg, about 17 Meq/kg, about 16 Meq/kg, about 15Meq/kg, about 14 Meq/kg, about 13 Meq/kg, about 12 Meq/kg, about 11Meq/kg, about 10 Meq/kg, about 9 Meq/kg, about 8 Meq/kg, about 7 Meq/kg,about 6 Meq/kg, about 5 Meq/kg, about 4 Meq/kg, about 3 Meq/kg or about2 Meq/kg.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising encapsulated EPA (e.g. E-EPA or ultra pure E-EPA)containing a labeled amount (i.e. initial amount) of EPA or E-EPA,wherein upon storage of the composition at 23° C. and 50% RH for aperiod about 1, about 2, about 3, about 4, about 5, about 6, about 7,about 8, about 9, about 10, about 11, about 12, about 13, about 14,about 15, about 16, about 17, about 18, about 19, about 20, about 21,about 22, about 23 or about 24 months, said composition contains atleast about 97%, about 98%, about 99%, about 99.5%, about 99.7%, about99.9% or substantially all or 100% of the labeled amount of EPA orE-EPA, by weight.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising encapsulated EPA (e.g. E-EPA or ultra pure E-EPA)containing a labeled amount (i.e. initial amount) of EPA or E-EPA,wherein upon storage of the composition at 25° C. and 60% RH for aperiod about 1, about 2, about 3, about 4, about 5, about 6, about 7,about 8, about 9, about 10, about 11, about 12, about 13, about 14,about 15, about 16, about 17, about 18, about 19, about 20, about 21,about 22, about 23 or about 24 months, said composition contains atleast about 97%, about 98%, about 99%, about 99.5%, about 99.7%, about99.9% or substantially all or 100% of the labeled amount of EPA orE-EPA, by weight.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising encapsulated EPA (e.g. E-EPA or ultra pure E-EPA)containing a labeled amount of EPA or E-EPA, wherein upon storage of thecomposition at 30° C. and 65% RH for a period about 1, about 2, about 3,about 4, about 5, about 6, about 7, about 8, about 9, about 10, about11, about 12, about 13, about 14, about 15, about 16, about 17, about18, about 19, about 20, about 21, about 22, about 23 or about 24 months,said composition contains at least about 97%, about 98%, about 99%,about 99.5%, about 99.7%, about 99.9%, substantially all or 100% of thelabeled amount of EPA or E-EPA, by weight.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising encapsulated EPA (e.g. E-EPA or ultra pure E-EPA)containing a labeled amount of EPA, wherein upon storage of thecomposition at 40° C. and 75% RH for a period about 1, about 2, about 3,about 4, about 5, about 6, about 7, about 8, about 9, about 10, about11, about 12, about 13, about 14, about 15, about 16, about 17, about18, about 19, about 20, about 21, about 22, about 23 or about 24 months,said composition contains at least about 97%, about 98%, about 99%,about 99.5%, about 99.7%, about 99.8%, about 99.9%, substantially all or100% of the labeled amount of EPA or E-EPA, by weight.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising encapsulated EPA containing a labeled amount ofEPA or E-EPA, wherein upon storage of the composition at 23° C. and 50%RH for a period about 1, about 2, about 3, about 4, about 5, about 6,about 7, about 8, about 9, about 10, about 11, about 12, about 13, about14, about 15, about 16, about 17, about 18, about 19, about 20, about21, about 22, about 23 or about 24 months, said composition contains notmore than about 0.5%, not more than about 0.25%, not more than about0.15%, not more than about 0.125%, not more than about 0.1%, not morethan about 0.075%, not more than about 0.05% or substantially nodegradation product and/or specified degradation product. The term“degradation product” in the present context means “an impurityresulting from a chemical change in the composition brought about duringmanufacture and/or storage of the composition by the effect of forexample, light, temperature, pH water or by reaction with an excipientand/or the immediate container closure system.” The term “specifieddegradation product in the present context means “a degradation product,either identified or unidentified, that is individually listed andlimited with a specific acceptance criterion in the productspecification” for a particular product.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising encapsulated EPA (e.g. E-EPA or ultra pure E-EPA)containing a labeled amount of EPA or E-EPA, wherein upon storage of thecomposition at 25° C. and 60% RH for a period about 1, about 2, about 3,about 4, about 5, about 6, about 7, about 8, about 9, about 10, about11, about 12, about 13, about 14, about 15, about 16, about 17, about18, about 19, about 20, about 21, about 22, about 23 or about 24 months,the composition contains not more than about 0.5%, not more than about0.25%, not more than about 0.15%, not more than about 0.125%, not morethan about 0.1%, not more than about 0.075%, not more than about 0.05%or substantially no degradation product and/or specified degradationproduct.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising encapsulated EPA (e.g. E-EPA or ultra pure E-EPA)containing a labeled amount of EPA or E-EPA, wherein upon storage of thecomposition at 30° C. and 65% RH or a period about 1, about 2, about 3,about 4, about 5, about 6, about 7, about 8, about 9, about 10, about11, about 12, about 13, about 14, about 15, about 16, about 17, about18, about 19, about 20, about 21, about 22, about 23 or about 24 months,the composition contains not more than about 0.5% (by weight of thelabeled EPA or E-EPA), not more than about 0.25%, not more than about0.15%, not more than about 0.125%, not more than about 0.1%, not morethan about 0.075%, not more than about 0.05% or substantially nodegradation product and/or specified degradation product.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising encapsulated EPA (e.g. E-EPA or ultra pure E-EPA)containing a labeled amount of EPA or E-EPA, wherein upon storage of thecomposition at 40° C. and 75% RH for a period about 1, about 2, about 3,about 4, about 5, about 6, about 7, about 8, about 9, about 10, about11, about 12, about 13, about 14, about 15, about 16, about 17, about18, about 19, about 20, about 21, about 22, about 23 or about 24 months,the composition contains not more than about 0.5% (by weight of thelabeled EPA or E-EPA), not more than about 0.25%, not more than about0.15%, not more than about 0.125%, not more than about 0.1%, not morethan about 0.075%, not more than about 0.05% or substantially nodegradation product and/or specified degradation product.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising encapsulated EPA (e.g. E-EPA or ultra pure E-EPA)containing a labeled amount of EPA or E-EPA, wherein the capsulecomprises a film-forming material, a hygroscopic plasticizer and anon-hygroscopic plasticizer and upon storage of the composition at 23°C. and 50% RH for a period about 1, about 2, about 3, about 4, about 5,about 6, about 7, about 8, about 9, about 10, about 11, about 12, about13, about 14, about 15, about 16, about 17, about 18, about 19, about20, about 21, about 22, about 23 or about 24 months, the compositioncontains not more than about 0.5% (by weight of the labeled EPA orE-EPA), not more than about 0.25%, not more than about 0.15%, not morethan about 0.125%, not more than about 0.1%, not more than about 0.075%,not more than about 0.05% or substantially no degradation product and/orspecified degradation product.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising encapsulated EPA (e.g. E-EPA or ultra pure E-EPA)containing a labeled amount of EPA, wherein the capsule comprises afilm-forming material, a hygroscopic plasticizer and a non-hygroscopicplasticizer and upon storage of the composition at 25° C. and 60% RH fora period about 1, about 2, about 3, about 4, about 5, about 6, about 7,about 8, about 9, about 10, about 11, about 12, about 13, about 14,about 15, about 16, about 17, about 18, about 19, about 20, about 21,about 22, about 23 or about 24 months, the composition contains not morethan about 0.5% (by weight of the labeled EPA or E-EPA), not more thanabout 0.25%, not more than about 0.15%, not more than about 0.125%, notmore than about 0.1%, not more than about 0.075%, not more than about0.05% or substantially no degradation product and/or specifieddegradation product.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising encapsulated EPA (e.g. E-EPA or ultra pure E-EPA)containing a labeled amount of EPA, wherein the capsule comprises afilm-forming material, a hygroscopic plasticizer and a non-hygroscopicplasticizer and upon storage of the composition at 30° C. and 65% RH fora period about 1, about 2, about 3, about 4, about 5, about 6, about 7,about 8, about 9, about 10, about 11, about 12, about 13, about 14,about 15, about 16, about 17, about 18, about 19, about 20, about 21,about 22, about 23 or about 24 months, the composition contains not morethan about 0.5% (by weight of the labeled EPA or E-EPA), not more thanabout 0.25%, not more than about 0.15%, not more than about 0.125%, notmore than about 0.1%, not more than about 0.075%, not more than about0.05% or substantially no degradation product and/or specifieddegradation product.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising EPA (e.g. E-EPA or ultra pure E-EPA) containing alabeled amount of EPA or E-EPA, wherein the capsule comprises afilm-forming material, a hygroscopic plasticizer and a non-hygroscopicplasticizer and upon storage of the composition at 40° C. and 75% RH fora period about 1, about 2, about 3, about 4, about 5, about 6, about 7,about 8, about 9, about 10, about 11, about 12, about 13, about 14,about 15, about 16, about 17, about 18, about 19, about 20, about 21,about 22, about 23 or about 24 months, the composition contains not morethan about 0.5% (by weight of the labeled EPA or E-EPA), not more thanabout 0.25%, not more than about 0.15% not more than about 0.125%, notmore than about 0.1%, not more than about 0.075%, not more than about0.05% or substantially no degradation product and/or specifieddegradation product.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising encapsulated EPA (e.g. E-EPA or ultra pure E-EPA)containing a labeled amount of EPA or E-EPA, wherein the capsulecomprises a film-forming material and a plasticizer in a weight ratio ofabout 2:5:1 to about 10:1 and upon storage of the composition at 23° C.and 50% RH for a period about 1, about 2, about 3, about 4, about 5,about 6, about 7, about 8, about 9, about 10, about 11, about 12, about13, about 14, about 15, about 16, about 17, about 18, about 19, about20, about 21, about 22, about 23 or about 24 months, the compositioncontains not more than about 0.5% (by weight of the labeled EPA orE-EPA), not more than about 0.25%, not more than about 0.15%, not morethan about 0.125%, not more than about 0.1%, not more than about 0.075%,not more than about 0.05% or substantially no degradation product and/orspecified degradation product.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising encapsulated EPA (e.g. E-EPA or ultra pure E-EPA)containing a labeled amount of EPA, wherein the capsule comprises afilm-forming material and a plasticizer in a weight ratio of about 2:5:1to about 10:1 and upon storage of the composition at 25° C. and 60% RHfor a period about 1, about 2, about 3, about 4, about 5, about 6, about7, about 8, about 9, about 10, about 11, about 12, about 13, about 14,about 15, about 16, about 17, about 18, about 19, about 20, about 21,about 22, about 23 or about 24 months, said composition contains notmore than about 0.5% (by weight of the labeled EPA or E-EPA), not morethan about 0.25, not more than about 0.15%, not more than about 0.125%,not more than about 0.1%, not more than about 0.075%, not more thanabout 0.05% or substantially no degradation product and/or specifieddegradation product.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising encapsulated (e.g. E-EPA or ultra pure E-EPA)containing a labeled amount of EPA or E-EPA, wherein the capsulecomprises a film-forming material and a plasticizer in a weight ratio ofabout 2:5:1 to about 10:1 and upon storage of the composition at 30° C.and 65% RH for a period about 1, about 2, about 3, about 4, about 5,about 6, about 7, about 8, about 9, about 10, about 11, about 12, about13, about 14, about 15, about 16, about 17, about 18, about 19, about20, about 21, about 22, about 23 or about 24 months, said compositioncontains not more than about 0.5% (by weight of the labeled EPA orE-EPA), not more than about 0.25%, not more than about 0.15%, not morethan about 0.125%, not more than about 0.1%, not more than about 0.075%,not more than about 0.05% or substantially no degradation product and/orspecified degradation product.

In another embodiment, the invention provides a pharmaceuticalcomposition comprising encapsulated (e.g. E-EPA or ultra pure E-EPA)containing a labeled amount of EPA or E-EPA, wherein the capsulecomprises a film-forming material and a plasticizer in a weight ratio ofabout 2:5:1 to about 10:1 and upon storage of the composition at 40° C.and 75% RH for a period about 1, about 2, about 3, about 4, about 5,about 6, about 7, about 8, about 9, about 10, about 11, about 12, about13, about 14, about 15, about 16, about 17, about 18, about 19, about20, about 21, about 22, about 23 or about 24 months, said compositioncontains not more than about 0.5% (by weight of the labeled EPA orE-EPA), not more than about 0.25%, not more than about 0.15%, not morethan about 0.125%, not more than about 0.1%, not more than about 0.075%,not more than about 0.05% or substantially no degradation product and/orspecified degradation product.

In another embodiment, the present invention provides a pharmaceuticalcomposition comprising about 0.5 g to about 1.5 g of EPA (e.g. E-EPA orultra pure E-EPA) having a labeled amount of EPA or E-EPA encapsulatedin a pharmaceutical capsule, wherein upon storage at 15° C. to 30° C.for a period of about 6 months, 12 months, 18 months, 24 months, 30months, or 36 months, at least about 97%, about 98%, about 99%, about99.5%, about 99.6%, about 99.7%, about 99.8%, about 99.9% orsubstantially all of the labeled amount of EPA is still present in thecomposition. In a related embodiment, the composition has not reachedits labeled expiration date during said storage period.

In various embodiments, capsule shells suitable for use in the presentinvention comprise one or more film-forming materials, one or moreplasticizers and optionally a solvent (e.g. water). In a relatedembodiment, the film-forming material comprises gelatin. In anotherembodiment, the plasticizer comprises a hygroscopic and/ornon-hygroscopic plasticizer. In still another embodiment, the capsuleshell comprises a film-forming material, a hygroscopic plasticizer, anon-hygroscopic plasticizer and a solvent.

In another embodiment, the capsule shell comprises about 30% to about70% or about 40% to about 65%, by weight, of a film-forming material,about 15% to about 40% or about 20% to about 35%, by weight, of one ormore plasticizers, and about 3% to about 15% or about 5% to about 10%,by weight, solvent such as water. Optionally, the capsules may alsocontain additives such as colorants, flavorants, preservatives,disintegrants, surfactants, fragrances, sweeteners, etc.

Capsules suitable for use in various embodiments of the inventioncomprise a film-forming material, for example gelatin. Gelatin istypically manufactured from animal byproducts that contain collagen, forexample in the bones, skin, and connective tissue. Methods of producinggelatin from animal byproducts are well-known in the art. In variousembodiments, the gelatin may be alkali-treated gelatin, acid-treatedgelatin, chemically modified gelatin, or mixtures thereof. Methods toproduce alkali-treated gelatin, acid-treated gelatin, and chemicallymodified gelatin are known in the art and are described, for example inNakamura et al., U.S. 2003/0195246, hereby incorporated by referenceherein in its entirety.

The film-forming material may also comprise, for example, non-animalbased hydrocolloids such as carrageenan, alkylated or hydroxyalkylatedcellulose ethers, starch, alpha-starch, hydroxyalkyl starch, sodiumalginate, sodium salt of a gelatin copolymer and acrylic acid.

In another embodiment, the film-forming material can comprise a 20:80 toabout 80:20, by weight, mixture, for example a 60:40, by weight mixtureof hydroxypropyl methyl cellulose and polyvinyl alcohol (e.g. about 70%to about 90%, for example about 88.0% saponified; and about 30 to about50, for example about 45.0 centipoise viscosity). In another embodiment,the film-forming material can comprise a 20:80 to about 80:20, byweight, mixture, for example a 60:40, by weight, mixture of hydroxyethylcellulose and polyvinyl alcohol (e.g. about 70% to about 99.9%, forexample about 98.5% saponified; and about 2 to about 30, for exampleabout 5.5 centipoise viscosity).

A suitable capsule shell may further comprise an elasticity reducing gelextender as part of the film-forming material. An elasticity reducinggel extender can comprise starch, starch derivatives such as highamylose starch, oxidized starch, esterified starch, acid-thinned starch,etherified starch, hydrolyzed starch, hydrolyzed and hydrogenatedstarch, enzyme treated starch, and modified celluloses or other naturalor modified natural biopolymers such as bacterial polysaccharides,vegetable gums, or other exudates including alginates, carrageenans,guar gum, gum arabic, gum ghatti, gum karaya, gum tragacanth, pectins,tamarind gum, xanthan gum, and dextrans as well as synthetic polymerssuch as carbon chain polymers of the vinyl and acrylic types as well asheterochains of the polyoxide and polyamine types including polyethyleneoxide, polypropylene oxide, polyoxymethylene, polytrimethylene oxide,block copolymers of ethylene oxide, block copolymers of polyethyleneoxide, polyvinyl methyl ether, polyethylene imine, polyacrylic acid,polyacrylamide, polymethacrylic acid, polymethacrylamide,poly(N,N-Dimethylacrylamide), poly(N-Isopropylacrylamide),poly(N-Acrylylglycinamide), poly(N-Methyacrylyglycinamide), acryliccopolymers, polyvinyl alcohol polyvinylacetate, polyvinylacetate-co-vinyl alcohol, polyvinylpyrrolidone, N-Methylpyrrolidone,N-Ethylpyrrolidone, N-Vinylpyrrolidone, sarcosine anhydride,polyvinyloxazolindone, and polyvinylmethyloxazolidone. The starch orother elasticity reducing gel extender may be added into the formulationin amounts ranging from about 8% to about 30% by weight, for exampleabout 10% to about 16%, by weight.

Capsule shells suitable for use in various embodiments of the inventioncan comprise one or more plasticizers, for example hygroscopic and/ornon-hygroscopic plasticizers. Non-limiting examples of suitablehygroscopic plasticizers include glycerin, sorbitol and alkylene glycols(e.g., propylene glycol and low molecular weight polyethylene glycols).Non-limiting examples of suitable non-hygroscopic plasticizers includepartially dehydrated hydrogenated glucose syrup, maltitol, maltose,lactitol, xylitol, erythritol and polyethylene glycols of averagemolecular weights from about 400 to about 6000.

In one embodiment, a capsule shell suitable for use in a composition ofthe invention has a hygroscopic plasticizer to non-hygroscopicplasticizer weight ratio of about 1:1 to about 8:1, about 2:1 to about6:1, about 3:1 to about 5:1, for example about 4:1, about 4.25:1, about4.5:1 or about 4.75:1.

In another embodiment, a capsule shell suitable for use in a compositionof the invention has a gelatin to glycerol weight ratio of about 2:5:1to about 10:1, about 3.5:1 to about 9:1, about 4:1 to about 8:1, orabout 5:1 to about 7:1, for example at least about 2.6:1, at least about2, 7:1, at least about 2.8:1, at least about 2.9:1, at least about 3:1,at least about 3.1:1, at least about 3.2:1, at least about 3.3:1, atleast about 3.4:1, at least about 3.5:1, at least about 3.6:1, at leastabout 3.7:1, at least about 3.8:1, at least about 3.9:1, at least about4.0:1, at least about 4.1:1, at least about 4.2:1, at least about 4.3:1,at least about 4.4:1, at least about 4.5:1, at least about 4.6:1, atleast about 4.7:1, at least about 4.8:1, at least about 4.9:1, at leastabout 5.0:1, at least about 5.1:1, or at least about 5.2:1.

In another embodiment, a suitable capsule shell has a film-formingmaterial (e.g. gelatin) to total plasticizer weight ratio of about 1.75to about 5, about 1.78 to about 3, or about 1.8 to about 2.5, forexample at least about 1.76, at least about 1.77, at least about 1.78,at least about 1.79, at least about 1.8, at least about 1.81, at leastabout 1.82, at least about 1.83, or at least about 1.84.

In another embodiment, the capsule shell has: (1) a gelatin to glycerolweight ratio of about 2:5:1 to about 10:1, about 3.5:1 to about 9:1,about 4:1 to about 8:1, or about 5:1 to about 7:1, for example at leastabout 2.6:1, at least about 2.7:1, at least about 2.8:1, at least about2.9:1, at least about 3:1, at least about 3.1:1, at least about 3.2:1,at least about 3.3:1, at least about 3.4:1, at least about 3.5:1, atleast about 3.6:1, at least about 3.7:1, at least about 3.8:1, at leastabout 3.9:1, at least about 4.0:1, at least about 4.1:1, at least about4.2:1, at least about 4.3:1, at least about 4, 4:1, at least about4.5:1, at least about 4.6:1, at least about 4.7:1, at least about 4.8:1,at least about 4.9:1, at least about 5.0:1, at least about 5.1:1, or atleast about 5.2:1; and/or (2) a gelatin to total plasticizer weightratio of about 1.75:1 to about 5:1, about 1.78:1 to about 3:1, or about1.8:1 to about 2.5:1, for example at least about 1.76:1, at least about1.77:1, at least about 1.78:1, at least about 1.79:1, at least about1.8:1, at least about 1.81, at least about 1.82, at least about 1.83, orat least about 1.84.

In one embodiment, the capsule shell comprises one or more of: gelatinin an amount of about 50% to about 70%; glycerol in an amount of about5% to about 15%; sorbitol in an amount of about 15% to about 25%; and/ormaltitol in an amount of about 3% to about 10%, by weight of thenon-aqueous components. Such a capsule can further comprise about 2% toabout 16% by weight of a solvent such as water.

In another embodiment, a capsule shell suitable for use in compositionsof the present invention can be prepared using a gel mass comprisingabout 40% to about 50% gelatin, about 2% to about 12% glycerol, about10% to about 20% sorbitol solution, about 2% to about 10% maltitolsyrup, and about 20% to about 35% water, by weight. In one embodiment, acapsule shell suitable for us in a composition of the present inventioncan be prepared using a gel mass comprising about 45% gelatin by weight,about 7% glycerol by weight, about 17% sorbitol solution (e.g. 30%water) by weight, about 6% maltitol syrup (e.g. 15%-32% water) byweight, and about 25% water by weight. Capsules prepared from such a gelmass can be dried to about 2% to about 12% final moisture content.Capsules prepared by such a process that contain EPA (e.g. E-EPA orultra pure E-EPA), and methods of using the same in the treatment ofcardiovascular-related diseases represent further embodiments of theinvention. Capsule compositions as described herein can further comprisecoatings, for example enteric polymer or wax coatings.

In one embodiment, a composition of the invention provides a relativelyrapid dissolution profile yet still maintains excellent stability of theencapsulated material (e.g. EPA). In a related embodiment, a compositionof the invention has a dissolution profile (as measured by RotatingDialysis Cell Dissolution (RDC) Apparatus under the conditions set forthherein below) of one or more of the following: (1) at least about 20%,at least about 23% or at least about 25% of E-EPA is dissolved by 10minutes; (2) at least about 45%, at least about 50% or at least about55° of E-EPA is dissolved by 30 minutes; (3) at least about 80%, atleast about 82%, at least about 85% or at least about 87% of E-EPA isdissolved by 60 minutes; and/or (4) at least about 95%, at least about97% or 100% of E-EPA is dissolved by 100 minutes. In a relatedembodiment, the fill material still retains the stability/peroxidevalues as set forth throughout this specification.

In another embodiment, a composition of the invention provides arelatively short T_(max) yet still maintains excellent stability of theencapsulated material (e.g. EPA). In a related embodiment, a compositionof the invention, upon administration to a subject, exhibits an EPAT_(max) less than 6 hours, less than 5.8 hours, less than 5.6 hours,less than 5.4 hours or less than 5.2 hours, for example about 4.8 toabout 5.2 hours. In a related embodiment, the fill material stillretains the stability/peroxide values as set forth throughout thisspecification.

In one embodiment, a method for treatment and/or prevention of acardiovascular-related disease using a composition as described hereinis provided. The term “cardiovascular-related disease” herein refers toany disease or disorder of the heart or blood vessels (i.e. arteries andveins) or any symptom thereof. The term “cardiovascular-related disease”herein refers to any disease or disorder of the heart or blood vessels(i.e. arteries and veins) or any symptom thereof or any disease orcondition that causes or contributes to a cardiovascular disease.”Non-limiting examples of cardiovascular-related diseases include acutecardiac ischemic events, acute myocardial infarction, angina, anginapectoris, arrhythmia, atrial fibrulation, atherosclerosis, arterialfibrillation, cardiac insufficiency, cardiovascular disease, chronicheart failure, chronic stable angina, congestive heart failure, coronaryartery disease, coronary heart disease, deep vein thrombosis, diabetes,diabetes mellitus, diabetic neuropathy, diastolic dysfunction insubjects with diabetes mellitus, edema, essential hypertension, eventualpulmonary embolism, fatty liver disease, heart disease, heart failure,homozygous familial hypercholesterolemia (HoFH), homozygous familialsitosterolemia, hypercholesterolemia, hyperlipidemia, hyperlipidemia inHIV positive subjects, hypertension, hypertriglyceridemia, ischemiccomplications in unstable angina and myocardial infarction, low bloodpressure, metabolic syndrome, mixed dyslipidemia, moderate to mild heartfailure, myocardial infarction, obesity management, paroxysmalatrial/arterial fibrillation/fibrulation/flutter, paroxysmalsupraventricular tachycardias (PSVT), particularly severe or rapid onsetedema, platelet aggregation, primary hypercholesterolemia, primaryhyperlipidemia, pulmonary arterial hypertension, pulmonary hypertension,recurrent hemodynamically unstable ventricular tachycardia (VT),recurrent ventricular arrhythmias, recurrent ventricular fibrillation(VF), ruptured aneurysm, sitisterolemia, stroke, supraventriculartachycardia, symptomatic atrial fibrillation/flutter, tachycardia,type-II diabetes, vascular disease, venous thromboembolism, ventriculararrhythmias, and other cardiovascular events.

The term “treatment” in relation a given disease or disorder, includes,but is not limited to, inhibiting the disease or disorder, for example,arresting the development of the disease or disorder; relieving thedisease or disorder, for example, causing regression of the disease ordisorder; or relieving a condition caused by or resulting from thedisease or disorder, for example, relieving, preventing or treatingsymptoms of the disease or disorder. The term “prevention” in relationto a given disease or disorder means: preventing the onset of diseasedevelopment if none had occurred, preventing the disease or disorderfrom occurring in a subject that may be predisposed to the disorder ordisease but has not yet been diagnosed as having the disorder ordisease, and/or preventing further disease/disorder development ifalready present.

In one embodiment, the present invention provides a method of bloodlipid therapy comprising administering to a subject or subject group inneed thereof a pharmaceutical composition as described herein. Inanother embodiment, the subject or subject group hashypertriglyceridemia, hypercholesterolemia, mixed dyslipidemia and/orvery high triglycerides.

In another embodiment, the subject or subject group being treated has abaseline triglyceride level (or median baseline triglyceride level inthe case of a subject group), fed or fasting, of at least about 300mg/dl, at least about 400 mg/dl, at least about 500 mg/dl, at leastabout 600 mg/dl, at least about 700 mg/dl, at least about 800 mg/dl, atleast about 900 mg/dl, at least about 1000 mg/dl, at least about 1100mg/dl, at least about 1200 mg/dl, at least about 1300 mg/dl, at leastabout 1400 mg/dl, or at least about 1500 mg/dl, for example about 400mg/dl to about 2500 mg/dl, about 450 mg/dl to about 2000 mg/dl or about500 mg/dl to about 1500 mg/dl.

In another embodiment, the subject or subject group being treated inaccordance with methods of the invention has previously been treatedwith Lovaza® and has experienced an increase in, or no decrease in,LDL-C levels and/or non-HDL-C levels. In one such embodiment, Lovaza®therapy is discontinued and replaced by a method of the presentinvention.

In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a fasting baselineabsolute plasma level of free EPA (or mean thereof in the case of asubject group) not greater than about 0.70 nmol/ml, not greater thanabout 0.65 nmol/ml, not greater than about 0.60 nmol/ml, not greaterthan about 0.55 nmol/ml, not greater than about 0.50 nmol/ml, notgreater than about 0.45 nmol/ml, or not greater than about 0.40 nmol/ml.In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a baseline fastingplasma level (or mean thereof) of free EPA, expressed as a percentage oftotal free fatty acid, of not more than about 3%, not more than about2.5%, not more than about 2%, not more than about 1.5%, not more thanabout 1%, not more than about 0.75%, not more than about 0.5%, not morethan about 0.25%, not more than about 0.2% or not more than about 0.15%.In one such embodiment, free plasma EPA and/or total fatty acid levelsare determined prior to initiating therapy.

In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a fasting baselineabsolute plasma level of total fatty acid (or mean thereof) not greaterthan about 250 nmol/ml, not greater than about 200 nmol/ml, not greaterthan about 150 nmol/ml, not greater than about 100 nmol/ml, or notgreater than about 50 nmol/ml.

In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a fasting baselineplasma, serum or red blood cell membrane EPA level not greater thanabout 70 μg/ml, not greater than about 60 μg/ml, not greater than about50 μg/ml, not greater than about 40 μg/ml, not greater than about 30μg/ml, or not greater than about 25 μg/ml.

In another embodiment, methods of the present invention comprise a stepof measuring the subject's (or subject group's mean) baseline lipidprofile prior to initiating therapy. In another embodiment, methods ofthe invention comprise the step of identifying a subject or subjectgroup having one or more of the following: baseline non-HDL-C value ofabout 200 mg/dl to about 400 mg/dl, for example at least about 210mg/dl, at least about 220 mg/dl, at least about 230 mg/dl, at leastabout 240 mg/dl, at least about 250 mg/dl, at least about 260 mg/dl, atleast about 270 mg/dl, at least about 280 mg/dl, at least about 290mg/dl, or at least about 300 mg/dl; baseline total cholesterol value ofabout 250 mg/dl to about 400 mg/dl, for example at least about 260mg/dl, at least about 270 mg/dl, at least about 280 mg/dl or at leastabout 290 mg/dl; baseline vLDL-C value of about 140 mg/dl to about 200mg/dl, for example at least about 150 mg/dl, at least about 160 mg/dl,at least about 170 mg/dl, at least about 180 mg/dl or at least about 190mg/dl; baseline HDL-C value of about 10 to about 60 mg/dl, for examplenot more than about 40 mg/dl, not more than about 35 mg/dl, not morethan about 30 mg/dl, not more than about 25 mg/dl, not more than about20 mg/dl, or not more than about 15 mg/dl; and/or baseline LDL-C valueof about 50 to about 300 mg/dl, for example not less than about 100mg/dl, not less than about 90 mg/dl, not less than about 80 mg/dl, notless than about 70 mg/dl, not less than about 60 mg/dl or not less thanabout 50 mg/dl.

In one embodiment, compositions of the invention are packaged in blisterpacks. In another embodiment, the blister packs comprise PCTFE (forexample 50μ) laminated with water based adhesive to clear PVC (forexample 190μ) which are heat sealed to aluminum foil).

In a related embodiment, upon treatment in accordance with the presentinvention, for example over a period of about 1 to about 200 weeks,about 1 to about 100 weeks, about 1 to about 80 weeks, about 1 to about50 weeks, about 1 to about 40 weeks, about 1 to about 20 weeks, about 1to about 15 weeks, about 1 to about 12 weeks, about 1 to about 10 weeks,about 1 to about 5 weeks, about 1 to about 2 weeks or about 1 week, thesubject or subject group exhibits one or more of the following outcomes:

(a) reduced triglyceride levels compared to baseline or a placebo arm;

(b) reduced Apo B levels compared to baseline or a placebo arm;

(c) increased HDL-C levels compared to baseline or a placebo arm;

(d) no increase in LDL-C levels compared to baseline or a placebo arm;

(e) a reduction in LDL-C levels compared to baseline or a placebo arm;

(f) a reduction in non-HDL-C levels compared to baseline or a placeboarm;

(g) a reduction in vLDL levels compared to baseline or a placebo arm;

(h) an increase in apo A-I levels compared to baseline or a placebo arm;

(i) an increase in apo A-I/apo B ratio compared to baseline or a placeboarm;

(j) a reduction in lipoprotein A levels compared to baseline or aplacebo arm;

(k) a reduction in LDL particle number compared to baseline or a placeboarm;

(l) an increase in mean LDL size compared to baseline or a placebo arm;

(m) a reduction in remnant-like particle cholesterol compared tobaseline or a placebo arm;

(n) a reduction in oxidized LDL compared to baseline or a placebo arm;

(o) no change or a reduction in fasting plasma glucose (FPG) compared tobaseline or a placebo arm;

(p) a reduction in hemoglobin A_(1c) (HbA_(1c)) compared to baseline ora placebo arm;

(q) a reduction in homeostasis model insulin resistance compared tobaseline or a placebo arm;

(r) a reduction in lipoprotein associated phospholipase A2 compared tobaseline or a placebo arm;

(s) a reduction in intracellular adhesion molecule-1 compared tobaseline or a placebo arm;

(t) a reduction in interleukin-6 compared to baseline or a placebo arm;

(u) a reduction in plasminogen activator inhibitor-1 compared tobaseline or a placebo arm;

(v) a reduction in high sensitivity C-reactive protein (hsCRP) comparedto baseline or a placebo arm;

(w) an increase in serum phospholipid EPA compared to baseline or aplacebo arm;

(x) an increase in red blood cell membrane EPA compared to baseline or aplacebo arm; and/or

(y) a reduction or increase in one or more of serum phospholipid and/orred blood cell content of docosahexaenoic acid (DHA), docosapentaenoicacid (DPA), arachidonic acid (AA), palmitic acid (PA), staeridonic acid(SA) or oleic acid (OA) compared to baseline or a placebo arm.

In one embodiment, methods of the present invention comprise measuringbaseline levels of one or more markers set forth in (a)-(y) above priorto dosing the subject or subject group. In another embodiment, themethods comprise administering a composition as disclosed herein to thesubject after baseline levels of one or more markers set forth in(a)-(y) are determined, and subsequently taking an additionalmeasurement of said one or more markers.

In another embodiment, upon treatment with a composition of the presentinvention, for example over a period of about 1 to about 200 weeks,about 1 to about 100 weeks, about 1 to about 80 weeks, about 1 to about50 weeks, about 1 to about 40 weeks, about 1 to about 20 weeks, about 1to about 15 weeks, about 1 to about 12 weeks, about 1 to about 10 weeks,about 1 to about 5 weeks, about 1 to about 2 weeks or about 1 week, thesubject or subject group exhibits any 2 or more of, any 3 or more of any4 or more of, any 5 or more of, any 6 or more of, any 7 or more of, any8 or more of, any 9 or more of, any 10 or more of, any 11 or more of,any 12 or more of, any 13 or more of, any 14 or more of, any 15 or moreof, any 16 or more of, any 17 or more of, any 18 or more of, any 19 ormore of, any 20 or more of, any 21 or more of, any 22 or more of, any 23or more, any 24 or more, or all 25 of outcomes (a)-(y) describedimmediately above.

In another embodiment, upon treatment with a composition of the presentinvention, the subject or subject group exhibits one or more of thefollowing outcomes:

(a) a reduction in triglyceride level of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) as compared to baseline or aplacebo arm;

(b) a less than 30% increase, less than 20% increase, less than 10%increase, less than 5% increase or no increase in non-H DL-C levels or areduction in non-HDL-C levels of at least about 1%, at least about 3%,at least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 55% or at least about 75%¹ (actual % change or median % change) ascompared to baseline or a placebo arm;

(c) substantially no change, no change or an increase in HDL-C levels ofat least about 5%, at least about 10) %, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 55% or at least about 75% (actual % change or median % change) ascompared to baseline or a placebo arm;

(d) a less than 60% increase, less than 50% increase, less than 40%increase, less than 30% increase, less than 20% increase, less than 10%increase, less than 5% increase or no increase in LDL-C levels or areduction in LDL-C levels of at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at about 45%, at leastabout 50%, at least about 55%, at least about 55% or at least about 75%(actual % change or median % change) as compared to baseline or aplacebo arm;

(e) a decrease in Apo B levels of at least about 5%, at least about 10%,at least about 15%, at least about 20%, at least about 25%, at leastabout 30%, at least about 35%, at least about 40%, at least about 45%,at least about 50%, at least about 55% or at least about 75% (actual %change or median % change) as compared to baseline or a placebo arm;

(f) a reduction in vLDL levels of at least about 5%, at least about 10%,at least about 15%, at least about 20%, at least about 25%, at leastabout 30%, at least about 35%, at least about 40%, at least about 45%,at least about 50%, or at least about 100% (actual % change or median %change) compared to baseline or a placebo arm;

(g) an increase in apo A-I levels of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, or at least about 100% (actual % change ormedian % change) compared to baseline or a placebo arm;

(h) an increase in apo A-I/apo B ratio of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30% at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, or at least about 100% (actual % changeor median % change) compared to baseline or a placebo arm;

(i) a reduction in lipoprotein(a) levels of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, or at least about 100% (actual % changeor median % change) compared to baseline or a placebo arm;

(j) a reduction in mean LDL particle number of at least about 5%, atleast about 10%, at least about 15%, at least about 20%, at least about25%, at least about 30%, at least about 35%, at least about 40%, atleast about 45%, at least about 50%, or at least about 100% (actual %change or median % change) compared to baseline or a placebo arm;

(k) an increase in mean LDL particle size of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, or at least about 100% (actual % changeor median % change) compared to baseline or a placebo arm;

(l) a reduction in remnant-like particle cholesterol of at least about5%, at least about 10%, at least about 15%, at least about 20%, at leastabout 25%, at least about 30%, at least about 35%, at least about 40%,at least about 45%, at least about 50%, or at least about 100% (actual %change or median % change) compared to baseline or a placebo arm;

(m) a reduction in oxidized LDL of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, or at least about 100% (actual % change ormedian % change) compared to baseline or a placebo arm;

(n) substantially no change, no change or a reduction in fasting plasmaglucose (FPG) of at least about 5%, at least about 10%, at least about15%, at least about 20%, at least about 25%, at least about 30%, atleast about 35%, at least about 40%, at least about 45%, at least about50%, or at least about 100% (actual % change or median % change)compared to baseline or a placebo arm;

(o) substantially no change, no change or a reduction in hemoglobinA_(1c) (HbA_(1c)) of at least about 5%, at least about 10%, at leastabout 15%, at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, or at leastabout 50% (actual % change or median % change) compared to baseline or aplacebo arm;

(p) a reduction in homeostasis model index insulin resistance of atleast about 5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 35%, atleast about 40%, at least about 45%, at least about 50%, or at leastabout 100% (actual % change or median % change) compared to baseline ora placebo arm;

(q) a reduction in lipoprotein associated phospholipase A2 of at leastabout 5%, at least about 10%, at least about 15%, at least about 20%, atleast about 25%, at least about 30%, at least about 35%, at least about40%, at least about 45%, at least about 50%, or at least about 100%(actual % change or median % change) compared to baseline or a placeboarm;

(r) a reduction in intracellular adhesion molecule-1 of at least about5%, at least about 10) %, at least about 15%, at least about 20%, atleast about 25%, at least about 30%, at least about 35%, at least about40%, at least about 45%, at least about 50%, or at least about 100%(actual % change or median % change) compared to baseline or a placeboarm;

(s) a reduction in interleukin-6 of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, or at least about 100% (actual % change ormedian % change) compared to baseline or a placebo arm;

(t) a reduction in plasminogen activator inhibitor-1 of at least about5%, at least about 10%, at least about 15%, at least about 20%, at leastabout 25%, at least about 30%, at least about 35%, at least about 40%,at least about 45%, at least about 50%, or at least about 100% (actual %change or median % change) compared to baseline;

(u) a reduction in high sensitivity C-reactive protein (hsCRP) of atleast about 5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 35%, atleast about 40%, at least about 45%, at least about 50%, or at leastabout 100% (actual % change or median % change) compared to baseline ora placebo arm;

(v) an increase in serum, plasma and/or RBC EPA of at least about 5%, atleast about 10%, at least about 15%, at least about 20%, at least about25%, at least about 30%, at least about 35%, at least about 40%, atleast about 45%, at least about 50%, at least about 100%, at least about200% or at least about 400% (actual % change or median % change)compared to baseline or a placebo arm;

(w) an increase in serum phospholipid and/or red blood cell membrane EPAof at least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, r at least about 50%, at leastabout 100%, at least about 200%, or at least about 400% (actual % changeor median % change) compared to baseline or a placebo arm;

(x) a reduction or increase in one or more of serum phospholipid and/orred blood cell DHA, DPA, AA, PA and/or OA of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) compared to baseline or a placeboarm; and/or

(y) a reduction in total cholesterol of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) compared to baseline or a placeboarm.

In one embodiment, methods of the present invention comprise measuringbaseline levels of one or more markers set forth in (a)-(y) prior todosing the subject or subject group. In another embodiment, the methodscomprise administering a composition as disclosed herein to the subjectafter baseline levels of one or more markers set forth in (a)-(y) aredetermined, and subsequently taking a second measurement of the one ormore markers as measured at baseline for comparison thereto.

In another embodiment, upon treatment with a composition of the presentinvention, for example over a period of about 1 to about 200 weeks,about 1 to about 100 weeks, about 1 to about 80 weeks, about 1 to about50 weeks, about 1 to about 40 weeks, about 1 to about 20 weeks, about 1to about 15 weeks, about 1 to about 12 weeks, about 1 to about 10 weeks,about 1 to about 5 weeks, about 1 to about 2 weeks or about 1 week, thesubject or subject group exhibits any 2 or more of, any 3 or more of,any 4 or more of, any 5 or more of, any 6 or more of, any 7 or more of,any 8 or more of, any 9 or more of, any 10 or more of, any 11 or moreof, any 12 or more of, any 13 or more of, any 14 or more of any 15 ormore of, any 16 or more of, any 17 or more of any 18 or more of, any 19or more of, any 20 or more of, any 21 or more of, any 22 or more of, any23 or more of, any 24 or more of, or all 25 of outcomes (a)-(y)described immediately above.

Parameters (a)-(y) can be measured in accordance with any clinicallyacceptable methodology. For example, triglycerides, total cholesterol,HDL-C and fasting blood sugar can be sample from serum and analyzedusing standard photometry techniques. VLDL-TG, LDL-C and VLDL-C can becalculated or determined using serum lipoprotein fractionation bypreparative ultracentrififugation and subsequent quantitative analysisby refractometry or by analytic ultracentrifugal methodology. Apo A1,Apo B and hsCRP can be determined from serum using standard nephelometrytechniques. Lipoprotein (a) can be determined from serum using standardturbidimetric immunoassay techniques. LDL particle number and particlesize can be determined using nuclear magnetic resonance (NMR)spectrometry. Remnants lipoproteins and LDL-phospholipase A2 can bedetermined from EDTA plasma or serum and serum, respectively, usingenzymatic immunoseparation techniques. Oxidized LDL, intercellularadhesion molecule-1 and interleukin-2 levels can be determined fromserum using standard enzyme immunoassay techniques. These techniques aredescribed in detail in standard textbooks, for example TietzFundamentals of Clinical Chemistry, 6^(th) Ed. (Burtis, Ashwood andBorter Eds.), WB Saunders Company.

In one embodiment, subjects fast for up to 12 hours prior to bloodsample collection, for example about 10 hours.

In another embodiment, the present invention provides a method oftreating or preventing primary hypercholesterolemia and/or mixeddyslipidemia (Fredrickson Types IIa and IIb) in a patient in needthereof, comprising administering to the patient one or morecompositions as disclosed herein. In a related embodiment, the presentinvention provides a method of reducing triglyceride levels in a subjector subjects when treatment with a statin or niacin extended-releasemonotherapy is considered inadequate (Frederickson type IVhyperlipidemia).

In another embodiment, the present invention provides a method oftreating or preventing risk of recurrent nonfatal myocardial infarctionin a patient with a history of myocardial infarction, comprisingadministering to the patient one or more compositions as disclosedherein.

In another embodiment, the present invention provides a method ofslowing progression of or promoting regression of atheroscleroticdisease in a patient in need thereof, comprising administering to asubject in need thereof one or more compositions as disclosed herein.

In another embodiment, the present invention provides a method oftreating or preventing very high serum triglyceride levels (e.g. TypesIV and V hyperlipidemia) in a patient in need thereof, comprisingadministering to the patient one or more compositions as disclosedherein.

In another embodiment, the present invention provides a method oftreating subjects having very high serum triglyceride levels (e.g.greater than 1000 mg/dl or greater than 2000 mg/dl) and that are at riskof developing pancreatitis, comprising administering to the patient oneor more compositions as disclosed herein.

In one embodiment, a composition of the invention is administered to asubject in an amount sufficient to provide a daily dose ofeicosapentaenoic acid of about 1 mg to about 10,000 mg, 25 about 5000mg, about 50 to about 3000 mg, about 75 mg to about 2500 mg, or about100 mg to about 1000 mg, for example about 75 mg, about 100 mg, about125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about375 mg, about 400 mg, about 425 mg, about 450 mg, about 475 mg, about500 mg, about 525 mg, about 550 mg, about 575 mg, about 600 mg, about625 mg, about 650 mg, about 675 mg, about 700 mg, about 725 mg, about750 mg, about 775 mg, about 800 mg, about 825 mg, about 850 mg, about875 mg, about 900 mg, about 925 mg, about 950 mg, about 975 mg, about1000 mg, about 1025 mg, about 1050 mg, about 1075 mg, about 1100 mg,about 1025 mg, about 1050 mg, about 1075 mg, about 1200 mg, about 1225mg, about 1250 mg, about 1275 mg, about 1300 mg, about 1325 mg, about1350 mg, about 1375 mg, about 1400 mg, about 1425 mg, about 1450 mg,about 1475 mg, about 1500 mg, about 1525 mg, about 1550 mg, about 1575mg, about 1600 mg, about 1625 mg, about 1650 mg, about 1675 mg, about1700 mg, about 1725 mg, about 1750 mg, about 1775 mg, about 1800 mg,about 1825 mg, about 1850 mg, about 1875 mg, about 1900 mg, about 1925mg, about 1950 mg, about 1975 mg, about 2000 mg, about 2025 mg, about2050 mg, about 2075 mg, about 2100 mg, about 2125 mg, about 2150 mg,about 2175 mg, about 2200 mg, about 2225 mg, about 2250 mg, about 2275mg, about 2300 mg, about 2325 mg, about 2350 mg, about 2375 mg, about2400 mg, about 2425 mg, about 2450 mg, about 2475 mg or about 2500 mg.

In another embodiment, any of the methods disclosed herein are used intreatment or prevention of a subject or subjects that consume atraditional Western diet. In one embodiment, the methods of theinvention include a step of identifying a subject as a Western dietconsumer or prudent diet consumer and then treating the subject if thesubject is deemed a Western diet consumer. The term “Western diet”herein refers generally to a typical diet consisting of, by percentageof total calories, about 45% to about 50% carbohydrate, about 35% toabout 40% fat, and about 10% to about 15% protein. A Western diet mayalternately or additionally be characterized by relatively high intakesof red and processed meats, sweets, refined grains, and desserts, forexample more than 50%, more than 60% or more or 70% of total caloriescome from these sources.

EXAMPLES

The following examples are for illustrative purposes only and should notbe construed as limiting the invention in any manner.

Example 1

A Test Composition (TC) was prepared comprising ultra-pure Ethyl-EPA(>96% E-EPA, ˜3% related fatty acid substances (no DHA), and ˜0.2% alphatocopherol) filled into soft gelatin capsule shells (˜500 mg fill weightper capsule) prepared from a gel comprising gelatin (˜44%), glycerol(˜7%), sorbitol solution (˜17%), maltitol solution, gelatin and purifiedwater. A Comparative Composition (CC) was made comprising the same fillas the Test Composition but filled into Type IIa Capsules made from agel comprising of glycerol (˜20%), gelatin (43.4%) and water (˜36.6%).

Test Compositions and Comparative Compositions were then placed inpolybags which were sealed and stored at either 25° C./60% RH or 30°C./65% RH for a period of 1, 3, or 6 months. At the end of storage,capsules were opened and peroxide value of the fill material wasanalyzed. Results are shown in Table 1 (average of capsules from threedifferent batches).

TABLE 1 Peroxide Values (Meq/kg) Upon Storage. Composition Baseline 1Month 3 Month 6 Month Storage at 25° C./60% RH TC 1.6 — 3.2 3.4 CC 1.9 —3.4 9.6 Storage at 30° C./65% RH TC 1.6 2.0 3.6 4.8 CC 1.8 1.9 3.5 12.5

As is seen in Table 1, the Test Composition fill material exhibited muchlower peroxide values after 6 months of storage under both sets ofstorage conditions. No significant differences were observed between theTest Composition and Comparative Composition fill material in terms ofpotency of EPA-E and related substances throughout the duration of thestudy.

Example 2

Test Compositions and Comparative Compositions of Example 1 wereprepared and packaged in blister packaging (50μ PCTFE laminated withwater based adhesive to 190μ clear PVC and heat sealed to aluminumfoil). Packaged Test Compositions and Comparative Compositions were thenstored at either 25° C./60% RH or 40° C./70% RH for a period of 1, 3, 6,12 or 36 months. At the end of storage, capsules were opened and theperoxide values of the fill contents analyzed as shown in Table 2(average of the three batches).

TABLE 2 Peroxide Values (Meq/kg) Upon Storage. Baseline 1 Mo. 3 Mo. 6Mo. 9 Mo. 12 Mo. Storage at 25° C./60% RH TC 2.5 — 1.1 2.1 2.2 5.4 CC2.6 — 5.1 8.3 9.7 11.1 Storage at 40° C./75% RH TC 2.5 2.1 3.2 4.9 — —CC 2.6 3.4 10.6 18.8 — —

As is seen in Table 2, the Test Composition exhibited much lowerperoxide values after 3, 6, 9 and 12 months of storage at 25° C./60% RHand after 1, 3 and 6 months of storage at 40° C./75% RH as compared tothe Comparative Compositions.

At 40° C., the Test Composition showed an average decrease in E-EPApotency of 0.30% per month whereas the Comparative Compositions showedan average decrease in E-EPA potency of 0.44% per month. However,similar results were not obtained with the same batches in Example 1(not stored in blister packages). Additionally, the related substancesmeasurements did not show any concomitant increase suggesting normalanalytic variation may be responsible.

When the peroxide values were forced to linear trendlines, average slopevalues between the Test Compositions in Experiment 1 (no blisterpackaging) and Experiment 2 (blister packaging) were similar indicatingthat the packaging is likely not responsible for prevention ofoxidation.

TABLE 3 Peroxide Value: Linear Slope Comparison Between Example 1 andExample 2 Test Composition Comparative Composition Storage Slope(Meq/kg/mo.) Slope (Meq/kg/mo.) Conditions Example 1 Example 2 Example 1Example 2 25° C./60% RH 0.33 0.35 1.45 1.03 40° C./75% RH 0.56 0.66 1.813.00

Example 3

A dissolution test was performed on capsules of Example 1 containing 500mg E-EPA using the Rotating Dialysis Cell method set forth in Yamazakiet al., Dissolution tests by RDC method for soft gelatin capsulescontaining ethyl icosapentate, Pharmaceutical Technology Japan, 15:595-603 (1999). Conditions were as set forth below:

-   -   RDC Cell: PharmaTest    -   Paddle speed: 100 rpm    -   Temperature: 37° C.    -   Filter: Millipore hydrophobic filter sheets    -   Inner media: JP pH 1.2 disintegration media    -   Outer Media Absolute Ethanol    -   Samples: 5 ml taken at 10, 20, 30, 40, 60, 100 and 120 minutes

The samples were analyzed against a reference standard prepared inethanol at 0.5 mg/ml, the amount of product dissolved at each time pointwas then calculated. A good dissolution profile was obtained with a Q₈₅of approximately 60 minutes and a profile very similar to that generatedby Yamazaki (JP data; succinated gelatin capsules). Dissolution profileof the inventive capsule composition was also evaluated by the paddlemethod in media containing buffer, SDS and IPA (100 rpm paddle speed,1000 ml, 37° C.). Samples were removed at intervals and analyzed againsta standard solution (9.5 ml/ml in methanol) by HPCL. All data are shownin FIG. 1.

Example 4

Bioavailability data were obtained for a capsule shell according toExample 1 containing 500 mg E-EPA (AMR101) and were compared againstreported by Yamazaki data for 300 mg Epadel capsules (succinatedgelatin; Comparitor 1 and Comparitor 2). Tmax data are shown in Table 4together with dissolution percentage at 60 min. Full bioavailabilityprofiles for EPA succinated capsules and AMR101 capsules are shown inFIGS. 2 and 3, respectively.

TABLE 4 Dissolution and Tmax. Dissolution at 60 min T_(max) (%) (hrs)Comparitor 11 77 6 Comparitor 2² 75 6 AMR101³ 87 5 ¹Capsule shell = 220mg; contents = 323 mg. ²Capsule shell = 134 mg; contents = 327 mg. ³Meanof three batches using RDC and pH 1.2.

As can be seen from Table 4, AMR101 exhibited greater E-EPA dissolutionby 60 minutes, and had a shorter T_(max) than was reported for Epadelpresent in succinated gelatin capsules.

1. A method of lowering triglycerides without substantially increasingLDL-C in a subject having fasting baseline triglycerides of 500 mg/dl toabout 1500 mg/dl comprising, administering to the subject about 4 g perday of a pharmaceutical composition comprising at least about 90%, byweight, ethyl eicosapentaenoate in one or more capsules comprising acapsule wall composed of a film-forming material that is free ofsuccinated gelatin for a period effective to reduce triglycerides by atleast 5% without substantially increasing LDL-C in the subject comparedto a second subject having fasting baseline triglycerides of 500 mg/dlto about 1500 mg/dl who has not received the pharmaceutical composition.2. The method of claim 1 wherein the film-forming material comprisesgelatin, carrageenan, alkylated or hydroxyalkylated cellulose ethers,starch, alpha-starch, hydroxyalkyl starch, sodium alginate, sodium saltof a gelatin copolymer and acrylic acid, hydroxypropyl methyl cellulose,hydroxyethyl cellulose or polyvinyl alcohol.
 3. The method of claim 1wherein the film-forming material comprises at least one plasticizer. 4.The method of claim 1 wherein the at least one plasticizer comprises ahygroscopic plasticizer.
 5. The method of claim 4 wherein thehygroscopic plasticizer comprises glycerol.
 6. The method of claim 1wherein the film-forming material comprises gelatin.
 7. The method ofclaim 1 comprising administering the composition to the subject for aperiod effective to reduce triglycerides by at least 15% withoutincreasing LDL-C compared to the second subject.
 8. The method of claim7 comprising administering the composition to the subject for a periodeffective to reduce triglycerides by at least about 25% withoutincreasing LDL-C compared to the second subject.
 9. The method of claim8 wherein said period is at least about 12 weeks.
 10. The method ofclaim 1 comprising administering the composition to the subject toeffect a reduction in apolipoprotein B compared to the second subject.11. The method of claim 1 comprising administering the composition tothe subject to effect at least a 5% reduction in apolipoprotein Bcompared to the second subject.